Chemokine receptor CXCR4 and its ligand CXCL12 expressions and clinical significance in bladder cancer.

It is well known that chemokine receptors and their ligands play important roles in mediating the invasion and metastasis of malignant tumors. This aim of this study was to investigate the expression and clinical significance of chemokine receptor CXCR4 and its ligand CXCL12 in bladder tumor tissues. Cancerous and adjacent normal bladder tissues were collected from 42 patients. The expressions of CXCR4 and CXCL12 proteins were then detected by immunohistochemistry, and the expressions of CXCR4 and CXCL12 mRNAs were detected by RT-PCR. Bladder cancer tissues showed higher positive expressions of CXCR4 and CXCL12 than those in normal bladder mucosal tissues (z = 7.332, 6.758, P < 0.001). Positive expressions of CXCR4 and CXCL12 were related to the differentiation degree and invasive depth of cancer tissues (z = 2.598-4.594, P < 0.05), but not to patient gender or age (z = 0.273-0.554, P > 0.05). The expression of CXCR4 was positively correlated to CXCL12 expression in bladder cancer tissues (r = 0.661, P < 0.05). RT-PCR revealed that CXCR4 and CXCL12 mRNAs were not expressed in normal tissues. Moreover, with increased depth of invasion, CXCR4 and CXCL12 mRNA expressions gradually increased in bladder cancer tissues and showed significant intergroup differences (F = 56.642, 67.928, P < 0.01). Taken together, these results indicate that the chemokine receptor CXCR4 and its ligand CXCL12 play important roles in the occurrence and development of bladder cancer.

Effect of siRNA targeting EZH2 on cell viability and apoptosis of bladder cancer T24 cells.

We investigated the effect of siRNA targeting enhancer of EZH2 on cell proliferation, invasion, migration, and apoptosis of human bladder cancer T24 cells. An siRNA-expressing plasmid targeting the EZH2 gene was transfected into T24 cells. Quantitative polymerase chain reaction and Western blot analysis were used to detect EZH2 expression at the mRNA and protein levels, respectively. Proliferation, invasion, and migration of T24 cells were examined in vivo using MTT, wound healing, and transwell chamber migration assays, respectively. Annexin V-fluorescein isothiocyanate/propidium iodide flow cytometric analysis was performed to determine cell apoptosis levels. Expression of EZH2 in T24 cells was suppressed at the mRNA and protein levels. Following transfection for 48 h, growth was inhibited by 37.9%, which was markedly lower than that in the negative control group (P < 0.05). Following a wound-healing assay for 24 h, transfected cell migration distance was 1.37 ± 0.12, which was markedly less than the horizontal migration distance of negative control group cells (P < 0.01). In addition, the cell invasion ability of EZH2- siRNA group cells decreased by 67% compared with negative control group cells (P < 0.01). Following transfection for 48 h, early- and late-stage apoptosis rates for T24 cells were 22.8 and 3.60%, respectively, which were higher than in the negative control group (P < 0.01). EZH2 gene silencing effectively suppressed the proliferation, invasion, and migration abilities of human bladder cancer cells, promoting apoptosis.

Assessment of the effects of prepartum anti-inflammatory therapies on type 1/type 2 immunity ratio using a rapid blood test.

The objective of this study was to assess (1) the effects of prepartum administration of anti-inflammatory therapies on type 1/type 2 immunity ratio using a rapid blood test (D2Dx immunity test; Nano Discovery Inc.), and (2) correlations between rapid blood test scores and daily milk yield in Holstein dairy cows. At 14 d before the expected calving date, cows (n = 64) and heifers (n = 23) were blocked by body condition score (optimal = 3.25-3.5; high ≥3.75) and parity (nulliparous, parous), and randomly allocated to one of 3 treatment groups (1) ASA (n = 29) = receive one oral treatment with administration of acetylsalicylic acid (4 boluses; 480 grain/bolus); (2) MEL (n = 31) = receive one oral administration with meloxicam (1 mg/kg of body weight), or (3) PLC (n = 27) = receive one oral treatment with 4 gelatin capsules filled with water. Blood samples were collected weekly starting 1 wk before treatment until 3 wk after calving for assessment of type 1/type 2 immunity ratio using a rapid blood test (i.e., D2Dx immunity test). A higher D2Dx score corresponds to a higher type 1/type 2 ratio. Furthermore, blood samples were collected within 72 h before and after calving by farm personnel. Daily milk yield for the first 60 d in milk (DIM) was collected from on-farm computer records. The data were analyzed using MIXED procedure of SAS (SAS Institute Inc.) as a randomized complete block design. On average enrolled cows received treatment administration 10 d before the actual calving date (standard deviation = 5.10 d). There was a tendency for a treatment by day interaction. Cows treated with ASA had higher type 1/type 2 ratio within 3 d after calving compared with MEL and PLC cows (ASA = 0.065 ± 0.002; MEL = 0.059 ± 0.002; PLC = 0.053 ± 0.002). Similarly, ASA and MEL cows had a higher type 1/type 2 ratio at 7 ± 3 DIM compared with PLC cows (ASA = 0.062 ± 0.002; MEL = 0.064 ± 0.002; PLC = 0.056 ± 0.002). Regardless of treatment, there was an interaction between parity and day. Parous cows had higher type 1/type 2 ratios compared with nulliparous cows at 14 ± 3 d before calving and at 7 ± 3, 14 ± 3, and 21 ± 3 d after calving. Furthermore, there was a positive correlation between D2Dx scores at 14 ± 3 DIM and average daily milk yield in the first 60 DIM. These results suggest that prepartum anti-inflammatory therapies may cause an increased shift in type 1 immunity around calving. Similarly, parous cows may have an increased shift in type 1 immunity after calving. Interestingly, higher type 1/type 2 ratios may be associated with higher milk yields in the first 60 DIM. Larger studies are needed to identify associations between the D2Dx immunity test and cow health and performance, as well as to assess the applicability of these types of tests in a conventional farm setting.

[Shikonin induces hepatocellular carcinoma cell apoptosis by suppressing PKM2/PHD3/HIF-1α signaling pathway].

OBJECTIVE: To investigate the mechanism of shikonin-induced death of human hepatocellular carcinoma SMMC-7721 cells. METHODS: Cultured SMMC-7721 cells and normal hepatocytes (L-02 cells) were treated with 4, 8, or 16 µmol/L shikonin, and the changes in cell viability was assessed using MTT assay. The levels of ATP and lactic acid in the cell cultures were detected using commercial kits. Co-immunoprecipitation and immunofluorescence staining were used to determine the relationship among pyruvate kinase M2 (PKM2), prolyl hydroxylase 3 (PHD3), and hypoxia-inducible factor-1α (HIF-1α). The expressions of PHD3, PKM2, HIF-1α, Bax, cleaved caspase-3, and Bcl-2 in SMMC-7721 cells were detected with Western blotting, and cell apoptosis was analyzed with annexin V-FITC/PI staining. The effects of RNA interference of PKM2 on PHD3 and HIF-1α expressions in SMMC-7721 cells were detected using Western blotting. RESULTS: The IC50 of shikonin against SMMC-7721 and L-02 cells was 8.041 µmol/L and 31.75 µmol/L, respectively. Treatment with shikonin significantly inhibited the protein expressions of PKM2, HIF-1α and PHD3 and nuclear translocation of PKM2 and HIF-1α in SMMC-7721 cells. Coimmunoprecipitation and immunofluorescence staining confirmed that shikonin inhibited the formation of PKM2/PHD3/HIF-1α complex and significantly reduced the contents of lactic acid and ATP in SMMC-7721 cells (P < 0.05). The expressions of PHD3 and HIF-1α decreased significantly after PKM2 knockdown (P < 0.05). Shikonin treatment significantly increased the apoptosis rate, enhanced the expressions of Bax and cleaved caspase-3, and decreased Bcl-2 expression in SMMC-7721 cells (P < 0.05). CONCLUSIONS: Shikonin induces apoptosis of SMMC-7721 cells possibly by inhibiting aerobic glycolysis through the PKM2/PHD3/HIF-1α signaling pathway to cause energy supply dysfunction in the cells.

Innocuous full-length botulinum neurotoxin targets and promotes the expression of lentiviral vectors in central and autonomic neurons.

Fragments of botulinum neurotoxin (BoNT) have been explored as potential targeting moieties and carriers of biomolecules into neurons, although with lower binding and translocation efficiency compared with intact proteins. This study exploits a detoxified recombinant form of full-length BoNT/B (BoTIM/B) fused with core streptavidin (CS-BoTIM/B) for lentiviral targeting to central and autonomic neurons. CS-BoTIM/B underwent an activity-dependent entry into cultured spinal cord neurons. Coupling CS-BoTIM/B to biotinylated lentivirus-encoding green fluorescent protein (GFP) endowed considerable neuron selectivity to the vector as evident from the preferential expression of the reporter in neurons co-cultured with skeletal muscle cells. CS-BoTIM/B-guided lentiviral transduction with the expression of a SNARE protein, SNAP-25 (S25), rendered non-susceptible to proteolysis by three BoNT serotypes, yielded a sizable decrease in cleaved S25 upon exposure of spinal cord neurons to these toxins. This was accompanied by synaptic transmission being spared from blockade by BoNT/A or BoNT/E, reflecting adequate translation and functional competence of recombinant multi-toxin-resistant S25. The augmented neurotropism conveyed on the lentivirus by CS-BoTIM/B was also demonstrated in vivo through enhanced expression of a reporter in intramural ganglionic neurons in the rat trachea, after injection of the targeted GFP-encoding lentivirus. Thus, a novel and realistic prospect for gene therapy of peripheral neuropathies is offered in this study through lentiviral targeting to neurons by CS-BoTIM/B.

Mesostructure design with gemini surfactants: supercage formation in a three-dimensional hexagonal array.

At low temperatures, liquid crystal-like arrays made up of inorganic-cluster and organic molecular units readily undergo reversible lyotropic transformations. Gemini surfactants, with two quaternary ammonium head groups separated by a methylene chain of variable length and with each head group attached to a hydrophobic tail, can be used to control organic charge sitting relative to the bivariable hydrophobic tail configurations. This approach has led to the synthesis of a mesophase (SBA-2) that has three-dimensional hexagonal (P6(3)/mmc) symmetry, regular supercages that can be dimensionally tailored, and a large inner surface area. This mesostructure analog of a zeolite cage structure does not appear to have a lyotropic surfactant or lipid liquid crystal mesophase counterpart. Through the modification of gemini charge separation and each of the two organic tails, these syntheses can be used to optimize templating effects, including the synthesis of MCM-48 at room temperature.

Cooperative organization of inorganic-surfactant and biomimetic assemblies.

A model that makes use of the cooperative organization of inorganic and organic molecular species into three dimensionally structured arrays is generalized for the synthesis of nanocomposite materials. In this model, the properties and structure of a system are determined by dynamic interplay among ion-pair inorganic and organic species, so that different phases can be readily obtained through small variations of controllable synthesis parameters, including mixture composition and temperature. Nucleation, growth, and phase transitions may be directed by the charge density, coordination, and steric requirements of the inorganic and organic species at the interface and not necessarily by a preformed structure. A specific example is presented in which organic molecules in the presence of multiply charged silicate oligomers self-assemble into silicatropic liquid crystals. The organization of these silicate-surfactant mesophases is investigated with and without interfacial silicate condensation to separate the effects of self-assembly from the kinetics of silicate polymerization.

Cooperative formation of inorganic-organic interfaces in the synthesis of silicate mesostructures.

A model is presented to explain the formation and morphologies of surfactant-silicate mesostructures. Three processes are identified: multidentate binding of silicate oligomers to the cationic surfactant, preferential silicate polymerization in the interface region, and charge density matching between the surfactant and the silicate. The model explains present experimental data, including the transformation between lamellar and hexagonal mesophases, and provides a guide for predicting conditions that favor the formation of lamellar, hexagonal, or cubic mesostructures. Model Q(230) proposed by Mariani and his co-workers satisfactorily fits the x-ray data collected on the cubic mesostructure material. This model suggests that the silicate polymer forms a unique infinite silicate sheet sitting on the gyroid minimal surface and separating the surfactant molecules into two disconnected volumes.

A new fluorescent chemosensor for copper ions based on tripeptide glycyl-histidyl-lysine (GHK).

[structure: see text]. A new fluorescent chemosensor for Cu2+ ions was synthesized by modifying the tripeptide glycyl-histidyl-lysine (GHK) with 9-carbonylanthracene via the standard Fmoc solid-phase peptide synthesis method. While significant fluorescence quenching was observed from the molecule upon binding with Cu2+, addition of Fe2+, Co2+, Ni2+, and Zn2+ to the peptide solution caused a minimum fluorescence emission spectral change, indicating a high specificity of this chemosensor for Cu2+ ions. Effects of pH were also investigated.

MicroRNA-30a suppresses breast tumor growth and metastasis by targeting metadherin.

Accumulating data have shown the involvement of microRNAs in cancerous processes as either oncogenes or tumor suppressor genes. Here, we established miR-30a as a tumor suppressor gene in breast cancer development and metastasis. Ectopic expression of miR-30a in breast cancer cell lines resulted in the suppression of cell growth and metastasis in vitro. Consistently, the xenograft mouse model also unveiled the suppressive effects of miR-30a on tumor growth and distal pulmonary metastasis. With dual luciferase reporter assay, we revealed that miR-30a could bind to the 3'-untranslated region of metadherin (MTDH) gene, thus exerting inhibitory effect on MTDH. Furthermore, we demonstrated that silence of MTDH could recapitulate the effects of miR-30a overexpression, while overexpression of MTDH could partially abrogate miR-30a-mediated suppression. Of significance, expression level of miR-30a was found to be significantly lower in primary breast cancer tissues than in the paired normal tissues. Further evaluation verified that miR-30a was negatively correlated with the extent of lymph node and lung metastasis in patients with breast cancer. Taken together, our findings indicated miR-30a inhibits breast cancer proliferation and metastasis by directly targeting MTDH, and miR-30a can serve as a prognostic marker for breast cancer. Manipulation of miR-30a may provide a promising therapeutic strategy for breast cancer treatment.

Microparticle association and heterogeneity of tumor-derived tissue factor in plasma: is it important for coagulation activation?

BACKGROUND: Tumor-derived tissue factor (TF) activates coagulation in vitro and in vivo in an orthotopic model of human pancreatic cancer. Here, we further characterized tumor-derived TF in this model. METHODS: Conditioned medium (CM) of L3.6pl human pancreatic tumor cells and plasma from nude mice bearing L3.6pl tumors were ultracentrifuged, and the pellets were filtered through membranes with different pore sizes. The size distribution of particles was analyzed in CM or plasma fractions with nanoparticle tracking and dynamic light scattering. Human TF antigen and activity were measured in pellets and supernatants with ELISA and clotting or thrombin generation assays, respectively. Human alternatively spliced TF (asTF) was measured with ELISA. Human TF and thrombin-antithrombin complex (TAT) concentrations were assessed in plasma of mice injected with filtered fractions of CM. RESULTS: Particles in both CM and plasma were < 0.4 µm. TF antigen and activity in the CM were mainly associated with microparticles (MP). Approximately 50% of antigen and 20% of activity were associated with particles of < 0.1 µm. Injection of < 0.1 µm particles into mice caused a 30% drop in platelet counts and an increase in TAT levels. In contrast, ~ 90% of TF antigen in tumor-bearing mice plasmas was non-sedimentable, whereas TF activity was exclusively associated with MP. Particles of < 0.1 µm and the supernatants of both CM and plasma gained TF activity after addition of exogenous phospholipids. Although asTF was found in MP-free CM supernatants, it was also present in CM and plasma pellets. CONCLUSIONS: Tumor-derived particles of < 0.1 µm and non-sedimentable TF are or can become procoagulant in the presence of phospholipids, and may contribute to the procoagulant potential of circulating TF.

Lyme borreliosis caused by diverse genospecies of Borrelia burgdorferi sensu lato in northeastern China.

The variety of Borrelia burgdorferi sensu lato (B. burgdorferi) genospecies leads to distinction in clinical manifestations of Lyme borreliosis (LB). There are reports of LB clinical characteristics in China, where the B. burgdorferi genospecies in ticks and animal hosts are different from those in Europe and North America. During May to September in 2010 and 2011, all patients who had erythema migrans (EM, more than 5 cm in diameter) after a recent tick-bite, and sought medical care at Mudanjiang Forestry Central Hospital, Heilongjiang Province of northeastern China, were enrolled in the study. Specific PCR was used to determine the B. burgdorferi genospecies in the disseminated patients. Of 265 EM patients, B. burgdorferi DNA was detected in blood specimens from 15 of 55 disseminated patients. Sequence analyses of 5S-23S rRNA, flagellin, ospC, 16S rRNA and ospA genes revealed that 11 patients were infected with Borrelia garinii, three with Borrelia afzelii and one with Borrelia valaisiana-related genospecies. Among 15 patients, 40%, 13.3% and 13.3% manifested pruritus, pain and ulceration, respectively. Systemic symptoms, arthralgia or a swollen joint and lymphadenopathy were observed in 26.7%, 13.3% and 6.7% patients, respectively. In northeastern China, three genospecies of LB patients were detected. The B. burgdorferi genospecies identified in this study was predominantly B. garinii. A case infected with B. valaisiana-related genospecies was reported for the first time.

The diversity of intestinal microbiota of Mongolians living in Inner Mongolia, China.

The Mongolian nationality has developed their unique lifestyle and dietary habit for thousands of years. However, by now, little research has been focused on Mongolian gut microbiota and how it is related to different dietary habits. In this study, denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction (qPCR) methods were applied to reveal the diversity of predominant gut bacteria of 48 healthy Mongolians recruited from Hohhot city and the Xilin Gol pasturing area in Inner Mongolia. Compared to similar studies of other nationalities, results from the present study have confirmed that the composition of Mongolian gut microbiota is highly similar at the phylum level (Firmicutes, Bacteroidetes, Proteobacteria and Actinobacteria) but variable at the genus level. Especially, the numbers of Phascolarctobacterium, Lactobacillus and Bifidobacterium are rather high. DGGE profiles of Lactobacillus and Bifidobacterium revealed that Lactobacillus casei, Bifidobacterium longum and Bifidobacterium animalis subsp. lactis were predominant in the gut of the Mongolian subjects studied. On the contrary, Lactobacillus helveticus was detected in every pasturing area Mongolian, but not in any of the Hohhot city Mongolians. qPCR results revealed that the numbers of Lactobacillus and Bifidobacterium of Xilin Gol Mongolians were significantly higher (P<0.05) than that of Hohhot Mongolians, whereas the numbers of Enterobacterium were significantly lower (P<0.05). In addition, by partial least squares discriminate analysis and cluster analysis of data generated from DGGE and qPCR experiments, a striking difference in the composition of intestinal microbiota of Mongolians living in Hohhot city and the Xilin Gol pasturing area has been found. This study clearly shows that diet affects the microbiota composition of Mongolians living in different circumstances, i.e. urban versus rural.

[The preparation of collagen burn pellicle of compound sulfadiazine silver and assessment of its efficacy in an animal experiment on deep partial thickness burn wound].

OBJECTIVE: To prepare the collagen burn pellicle of compound sulfadiazine sliver and observe its therapeutic effect on deep partial thickness burn wound. METHODS: The initator method was adopted for preparing the collagen burn pellicle of compound sulfadiazine sliver. A model of deep partial thickness burn wound was established in 84 SD rats for the observation of the effect of collagen burn pellicle of compound sulfadiazine sliver on burn wound healing. RESULTS: The collagen burn pellicle of compound sulfadiazine sliver enhanced the coming off of necrotic tissues and the healing of burn wound. The hydroxyproline content of burn wound was higher in the experiment group than that in the comparison groups, (P < 0.05). The percentage of G0/G1-phase in full skin cells of burn wound at 5, 7, 10 and 14 days after burn was lower than that in the comparison groups (P < 0.05). The percentage of S-phase at 5, 7 and 14 days was higher in the experiment group than that in the comparison groups (P < 0.05). The water content in full skin cells of burn wound at 24, 36 and 48 hours after burn was significantly lower in the experiment group than that the comparison groups (P < 0.05). CONCLUSION: The collagen burn pellicle of compound sulfadiazine sliver can enhance wound healing in the management of deep partial thickness burn wound.

The transport kinetics of lanthanide species in a single erythrocyte probed by confocal laser scanning microscopy.

A novel method has been developed to visualize and follow the temporal course of lanthanide transport across the membrane into a single living erythrocyte. By means of confocal scanning microscopy and the optical section technique, the entry of lanthanide ions was followed by the fluorescence quenching of fluorescein isothiocyanate (FITC)-labeled membrane and cytosol. From the difference of the quenching kinetics of the whole section and the central area, the time for diffusion through the membrane and the diffusion in the extracellular and intracellular media can be deduced. To clarify the mechanism of lanthanide-induced fluorescence quenching of FITC-labeled erythrocytes and to ensure that this reaction can be used in this method, the reaction was investigated by steady-state fluorescence techniques. The results showed that the lanthanides strongly quenched the florescence emitted by FITC covalently bound to membrane proteins and cytosolic proteins. The static quenching mechanism is responsible for the fluorescence quenching of FITC-labeled proteins by Ln species. The quenching mechanism is discussed on the basis of complex formation. The dependence of fluorescence quenching on both ion size and the total orbital angular momentum L supports the complexation mechanism. The transport time across the membrane is strikingly correlated with Ln species and extracellular concentration. For a given concentration, the transport time of [Ln(cit)2]3- is much shorter than that of Ln3+, since they enter the cells via the anion channel. This is supported by the inhibition effect of 4,4'-diisothiocyanato-2,2'-stilbenendisulfonate on the transport of [Ln(cit)2]3-. On the other hand, the transport of free Ln3+ might be attributed to the enhanced permeability of erythrocytes owing to Ln3+ binding. These findings strongly demonstrate the existence of the non-internalization mechanism of Ln species uptake by erythrocytes.

Metal complexation with Langmuir monolayers of histidyl peptide lipids.

Langmuir monolayers made from peptide-lipid molecules represent a novel direction in the research areas of biomimetic interfaces and two-dimensional supramolecular chemistry. Peptide structures and molecular recognition activities toward other guest molecules have been the focus of previous study. This study reports the investigation of metal complexation to histidine-containing peptide lipids in the organized Langmuir, Langmuir-Schaefer, or Langmuir-Blodgett films. Three peptide lipids PEP1-PEP3, with a histidine amino acid incorporated in the middle of the peptide, were designed and synthesized. The monolayer structures and metal-binding activities of each peptide lipid and their 1:1:1 molar ratio mixture were studied by thermodynamic and spectroscopic techniques. It was found that hard Lewis acid type metal cations such as K+ and Mg2+, and borderline or soft metal cations such as Zn2+, Cu2+, and Cd2+ exhibit clearly different binding activity toward peptide-lipid monolayers. The conformational changes of peptides upon binding with Cu2+ and Zn2+ were partially revealed by FT-IR spectroscopic studies. Furthermore, by adding a fluorescent-probe lipid to the peptide monolayer, dramatic fluorescence change was observed when Cu2+ or Zn2+ bound to the Langmuir and Langmuir-Schaefer films of peptide-lipid monolayers. Metal-protein complexation plays a crucial role in the function and activity of proteins and enzymes. Investigation of metal complexation to organized peptide Langmuir monolayers may provide an alternative approach for the development of artificial metalloproteins and novel supramolecular systems or materials.

Morphine inhibited respiratory burst of neutrophils and scavenged oxygen free radicals.

AIM: To study the effects of morphine on active oxygen free radicals. METHOD: Chemiluminescence method was used to measure (a) active oxygen generation induced by respiratory burst of neutrophils from human blood stimulated with phorbol myristate acetate (PMA), (b) Superoxide anion (O2.-) induced by xanthine-xanthine oxidase system, (c) hydroxyl radical (.OH) generated by ascorbic acid (AA)-Cu(2+)-zymosan, and (d) the release of H2O2. RESULTS: The (a), (b), (c), and (d) were scavenged by morphine and their median inhibitory concentrations (IC50 and 95% confidence limits) were 21.1 (13.0-34.0), 54.1 (50.0-58.5), 224.0 (128.2-390.8), and 66.9 (62.9-71.0), nmol L-1, respectively. CONCLUSION: The immunosuppressant effect of morphine was related to its free radicals scavenging action.

Oil-Water Interface Templating of Mesoporous Macroscale Structures

Ordered mesostructured porous silicas that are also macroscopically structured were created by control of the interface on two different length scales simultaneously. Micellar arrays controlled the nanometer-scale assembly, and at the static boundary between an aqueous phase and an organic phase, control was achieved on the micrometer to centimeter scale. Acid-prepared mesostructures of silica were made with the p6, Pm3n, and the P63/mmc structures in the form of porous fibers 50 to 1000 micrometers in length, hollow spheres with diameters of 1 to 100 micrometers, and thin sheets up to 10 centimeters in diameter and about 10 to 500 micrometers in thickness. These results might have implications for technical applications, such as slow drug-release systems or membranes, and in biomineralization, where many processes are also interface-controlled.